溴莫尼定治疗大鼠视神经夹挫伤机制探讨
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摘要
目的:外伤性视神经病变(Traumatic Optic Neuropathy , TON)是指外力通过骨质或眼球的移动传递给视神经造成的间接损伤,是眼科常见致盲性眼病之一,这种损伤在闭合性脑部外伤中约占0. 5 %～5 %[1、2]。间接性外伤性视神经病变引起的视力损伤的程度各不相同,约50%的患者仅为“光感”或“无光感”视力。目前还没有确凿的治疗方案。近年来随着对细胞凋亡现象认识的深入及国内外大量视神经损伤与修复方面的研究,神经不可再生的观点越来越引起医学界质疑,神经再生问题的进一步研究有望引发医学研究领域的又一重大突破。国内外大量研究显示药物、手术、基因治疗等多种治疗方法均显示对视网膜神经节细胞(Retinal Ganglion Cells ,RGCs)的再生有一定促进作用。笔者通过大量查阅国内外权威文献,了解到视神经损伤后轴突的再生是一个多环节、多因素的过程,而α2-肾上腺素受体激动剂(α2-Selective Adrenergic Agonists ,α2-SAs)在动物实验中已被证实有确切疗效,但其具体作用机制仍不清楚,本课题通过制作大鼠视神经夹挫伤动物模型,应用α2-SAs(0.2%酒石酸溴莫尼定)进行干预,并通过免疫组织化学检测大鼠视网膜中bFGF、Bcl-2、Bax的表达,用透射电镜观察微观形态,以探讨该药在治疗视神经夹挫伤中的具体作用机制,为临床更好地治疗视神经损伤提供新的思路和策略,并增进神经再生问题方面的认识。
方法
1 90只雌性S-D大鼠随机分为正常组、未治疗组和治疗组,未治疗组和治疗组采用反向镊钳夹法制作大鼠视神经夹挫伤动物模型,其中治疗组大鼠在造模前及造模后给予0.2%酒石酸溴莫尼定滴眼液点眼,未治疗组及正常组不做任何处理。
2分别于造模后l、3、5、7、14、21d通过氯胺酮肌肉注射麻醉未治疗组、治疗组大鼠及相应时间点正常组大鼠,摘出眼球固定于4%多聚甲醛溶液中,沿眼球赤道部剪开,弃去眼前节及眼内容物,取颞上象限距视神经2mm以内的视网膜组织切片并行HE染色,光镜下进行RGCs计数以观察RGCs的动态变化。
3采用同样方法麻醉各组不同时间点用作免疫组织化学染色的大鼠,摘出眼球固定于4%多聚甲醛溶液中,取颞上象限距视神经2mm以内的视网膜组织行bFGF、Bcl-2、bax免疫组织化学染色,观察三种因子在各组大鼠视网膜中不同时间点的表达及差异。
4分别于3、7、21d麻醉用于透射电镜观察的三组大鼠,迅速摘出眼球保存于2.5%戊二醛保存液中,取视神经周围部位视网膜组织送做透射电镜,观察三组大鼠视网膜不同时期的微观形态变化及差异。
结果
1光镜及透射电镜下观察到正常组大鼠视网膜各层细胞排列整齐致密,未治疗组视网膜各层细胞排列不规则,总厚度变薄,出现明显凋亡形态变化,治疗组形态变化较未治疗组有明显减轻。
2 Bcl-2在正常组大鼠视网膜中的阳性反应主要出现于视神经节细胞层,未治疗组损伤后随损伤时间的延长,Bcl-2在视网膜中的表达逐渐增强,损伤后第7、14、21d时,呈强阳性反应;治疗组较未治疗组表达明显增加,有统计学差异(p<0.01)。
3 Bax在正常组大鼠视网膜中有表达,阳性反应主要出现于神经节细胞层、内丛状层、内核层,未治疗组随视神经损伤时间的延长,Bax在3、5、7d未治疗组大鼠视网膜中的表达强度增强,且治疗组表达强度明显小于未治疗组,有统计学差异(p<0.05)。
4 bFGF在正常组大鼠视网膜中主要分布于视神经节细胞层及内核层,未治疗组视神经节细胞层及内核层染色强度增加,外核层出现bFGF阳性细胞;治疗组较未治疗组阳性表达有明显增强,有统计学差异(p<0.05)。
结论
1 0.2%酒石酸溴莫尼定滴眼液明显减轻大鼠视神经夹挫伤后视网膜形态损伤变化,对大鼠视神经夹挫伤有较好的疗效。
2 0.2%酒石酸溴莫尼定滴眼液点眼使视神经夹挫伤大鼠视网膜中bFGF表达明显增加;Bcl-2表达增加,Bax表达减少,Bcl-2/Bax比率明显增大,提示酒石酸溴莫尼定治疗大鼠视神经夹挫伤的作用机制可能与促进bFGF表达增加和抑制凋亡有关。
Objective: Traumatic optic neuropathy (TON) is an common ophthalmopathy that can leading to blind which can be indirectly caused by the extra-power passing from bone or eye ball. This kind of injury occupies 0.5%～5% in the closed brain trauma[1、2]. Degrees of the visual acuity damage indirectly caused by the TON differ from each other, however, about 50% of them are“light perception”or“non-light perception”. There is still no surely useful treatment plan for treating this kind of disease at present. With the development of the research on cell apoptosis and the optic nerve trauma and renovation, the view that nerve can not regeneration evoks more and more doubts in medical field. The continuing of research on the nerve regenation may initiates another breakthrough in medical domain.Many researches in our nation and overseas indicate that lots of new treatment methods show invariably effection in promoting the regeneration of the RGCs, such as medicine therapy, operation therapy and gene therapy.Author refers to lots of authority documents in domestic or overseas, and understands that the regeneration of the axonal after the optic nerve trauma is an complicated process including multiple factors and elements, andα2-selective adrenergic agonists (α2-SAs) has been proved to have evident effection in animal experiment, but its concrete mechanism of action is still unknown. This topic aims to research the concrete mechanism of action of the 0.2%brimonidine tartrate in therapying the optic nerve crush injury rat from immunohistochemistry of the Bcl-2, Bax, bFGF and the transmission electron microscope (TEM). We expect that this research may provide us some new strategies for treating optic nerve trauma and expand our recognition on nerve regeneration.
Methods
1 90 female adult Sprague-Dawley rats were randomly divided into normal group, untreated group and treated group. The untreated and treated groups received an optic nerve crush injury by reverse pliers; Then the treated group was gieven drops of 0.2%brimonidine tartrate topically before and after the crush injury;The untreated and normal groups received none intervention.
2 All three groups were anesthetized with ketamine by 1, 3, 5, 7, 14, 21d after the optic nerve crush injury,and all the eye balls were enucleate and fixed into 4% paraformaldehyde for 72 hours,then the eye balls were sheared along the ambitus ,eye protomerite and eye content were removed.The part of temple upper quadrant rat retina which leave optic papillary within 2mm was selected to cutting sheet and HE dyeing,then RGCs number were counted out under the light microscope.
3 Rats for immunohistochemistry were anesthetized with the same method , and all the eye balls were enucleate and fixed into 4% paraformaldehyde ,then the part of temple upper quadrant rat retina that leave the optic papillary within 2mm was selected for Bcl-2,Bax,bFGF immunohistochemistry staining.
4 Rats from three groups for TEM were anesthetized by 3,7,21d,and eye balls were enucleate quickly and fixed into 2.5% glutaraldehyde. Retina tissue around the optic papillary was selected for TEM in order to observe the microcosmic morphologic change among three groups.
Results
1 Bax expression was observed in normal group rats retina and its potitive staining mainly located at ganglion cell layer, inner plexiform layer,inner nuclear layer.The expression of Bax was gradually strengthened by 3, 5, 7d in untreated group.The expression of Bax in untreated group is higher than that in treated group,and the statistics difference is significant (p<0.05) .
2 The positive expression of Bcl-2 in normal group rat retina was mainly located at ganglion cell layer, The expression of Bcl-2 was gradually strengthened in untreated group and showed strong positive action by 7, 14, 21d. The expression of Bcl-2 in treated group is higher than that in untreated group,and the statistics difference is significant (p<0.01) .
3 The positive expression of bFGF in normal group rat retina was mainly located at ganglion cell layer and inner nuclear layer.The staining in ganglion cell layer and inner nuclear layer were increased in untreated group ,and positive cells were observed in outer nuclear layer. The expression of bFGF in treated group is higher than that in untreated group,and the statistics difference is significant (p<0.01) .
4 Every sheet of cells in normal group rats retina lined up in order under the light microscope and TEM;Cells in untreated group lined up irregularly,the whole thickness thinningzed,and manifest apoptosis morphologic change was observed.Mohphologic change in treated group was lessened obviously.
Conclusions
1 0.2%brimonidine tartrate relieves the apoptosis morphologic change and elevate survival rate of RGCs, showing obvious effect in threapying rat optic nerve crush injury.
2 0.2% brimonidine tartrate elevates the expression of bFGF、Bcl-2 ,cuts down the expression of the Bax in optic nerve crush injury rat retina,this hints that the therapeutic mechanism of 0.2% brimonidine tartrate may relevants with its effects of promoting the expression of bFGF and inhibiting apoptosis.
方法
1 90只雌性S-D大鼠随机分为正常组、未治疗组和治疗组,未治疗组和治疗组采用反向镊钳夹法制作大鼠视神经夹挫伤动物模型,其中治疗组大鼠在造模前及造模后给予0.2%酒石酸溴莫尼定滴眼液点眼,未治疗组及正常组不做任何处理。
2分别于造模后l、3、5、7、14、21d通过氯胺酮肌肉注射麻醉未治疗组、治疗组大鼠及相应时间点正常组大鼠,摘出眼球固定于4%多聚甲醛溶液中,沿眼球赤道部剪开,弃去眼前节及眼内容物,取颞上象限距视神经2mm以内的视网膜组织切片并行HE染色,光镜下进行RGCs计数以观察RGCs的动态变化。
3采用同样方法麻醉各组不同时间点用作免疫组织化学染色的大鼠,摘出眼球固定于4%多聚甲醛溶液中,取颞上象限距视神经2mm以内的视网膜组织行bFGF、Bcl-2、bax免疫组织化学染色,观察三种因子在各组大鼠视网膜中不同时间点的表达及差异。
4分别于3、7、21d麻醉用于透射电镜观察的三组大鼠,迅速摘出眼球保存于2.5%戊二醛保存液中,取视神经周围部位视网膜组织送做透射电镜,观察三组大鼠视网膜不同时期的微观形态变化及差异。
结果
1光镜及透射电镜下观察到正常组大鼠视网膜各层细胞排列整齐致密,未治疗组视网膜各层细胞排列不规则,总厚度变薄,出现明显凋亡形态变化,治疗组形态变化较未治疗组有明显减轻。
2 Bcl-2在正常组大鼠视网膜中的阳性反应主要出现于视神经节细胞层,未治疗组损伤后随损伤时间的延长,Bcl-2在视网膜中的表达逐渐增强,损伤后第7、14、21d时,呈强阳性反应;治疗组较未治疗组表达明显增加,有统计学差异(p<0.01)。
3 Bax在正常组大鼠视网膜中有表达,阳性反应主要出现于神经节细胞层、内丛状层、内核层,未治疗组随视神经损伤时间的延长,Bax在3、5、7d未治疗组大鼠视网膜中的表达强度增强,且治疗组表达强度明显小于未治疗组,有统计学差异(p<0.05)。
4 bFGF在正常组大鼠视网膜中主要分布于视神经节细胞层及内核层,未治疗组视神经节细胞层及内核层染色强度增加,外核层出现bFGF阳性细胞;治疗组较未治疗组阳性表达有明显增强,有统计学差异(p<0.05)。
结论
1 0.2%酒石酸溴莫尼定滴眼液明显减轻大鼠视神经夹挫伤后视网膜形态损伤变化,对大鼠视神经夹挫伤有较好的疗效。
2 0.2%酒石酸溴莫尼定滴眼液点眼使视神经夹挫伤大鼠视网膜中bFGF表达明显增加;Bcl-2表达增加,Bax表达减少,Bcl-2/Bax比率明显增大,提示酒石酸溴莫尼定治疗大鼠视神经夹挫伤的作用机制可能与促进bFGF表达增加和抑制凋亡有关。
Objective: Traumatic optic neuropathy (TON) is an common ophthalmopathy that can leading to blind which can be indirectly caused by the extra-power passing from bone or eye ball. This kind of injury occupies 0.5%～5% in the closed brain trauma[1、2]. Degrees of the visual acuity damage indirectly caused by the TON differ from each other, however, about 50% of them are“light perception”or“non-light perception”. There is still no surely useful treatment plan for treating this kind of disease at present. With the development of the research on cell apoptosis and the optic nerve trauma and renovation, the view that nerve can not regeneration evoks more and more doubts in medical field. The continuing of research on the nerve regenation may initiates another breakthrough in medical domain.Many researches in our nation and overseas indicate that lots of new treatment methods show invariably effection in promoting the regeneration of the RGCs, such as medicine therapy, operation therapy and gene therapy.Author refers to lots of authority documents in domestic or overseas, and understands that the regeneration of the axonal after the optic nerve trauma is an complicated process including multiple factors and elements, andα2-selective adrenergic agonists (α2-SAs) has been proved to have evident effection in animal experiment, but its concrete mechanism of action is still unknown. This topic aims to research the concrete mechanism of action of the 0.2%brimonidine tartrate in therapying the optic nerve crush injury rat from immunohistochemistry of the Bcl-2, Bax, bFGF and the transmission electron microscope (TEM). We expect that this research may provide us some new strategies for treating optic nerve trauma and expand our recognition on nerve regeneration.
Methods
1 90 female adult Sprague-Dawley rats were randomly divided into normal group, untreated group and treated group. The untreated and treated groups received an optic nerve crush injury by reverse pliers; Then the treated group was gieven drops of 0.2%brimonidine tartrate topically before and after the crush injury;The untreated and normal groups received none intervention.
2 All three groups were anesthetized with ketamine by 1, 3, 5, 7, 14, 21d after the optic nerve crush injury,and all the eye balls were enucleate and fixed into 4% paraformaldehyde for 72 hours,then the eye balls were sheared along the ambitus ,eye protomerite and eye content were removed.The part of temple upper quadrant rat retina which leave optic papillary within 2mm was selected to cutting sheet and HE dyeing,then RGCs number were counted out under the light microscope.
3 Rats for immunohistochemistry were anesthetized with the same method , and all the eye balls were enucleate and fixed into 4% paraformaldehyde ,then the part of temple upper quadrant rat retina that leave the optic papillary within 2mm was selected for Bcl-2,Bax,bFGF immunohistochemistry staining.
4 Rats from three groups for TEM were anesthetized by 3,7,21d,and eye balls were enucleate quickly and fixed into 2.5% glutaraldehyde. Retina tissue around the optic papillary was selected for TEM in order to observe the microcosmic morphologic change among three groups.
Results
1 Bax expression was observed in normal group rats retina and its potitive staining mainly located at ganglion cell layer, inner plexiform layer,inner nuclear layer.The expression of Bax was gradually strengthened by 3, 5, 7d in untreated group.The expression of Bax in untreated group is higher than that in treated group,and the statistics difference is significant (p<0.05) .
2 The positive expression of Bcl-2 in normal group rat retina was mainly located at ganglion cell layer, The expression of Bcl-2 was gradually strengthened in untreated group and showed strong positive action by 7, 14, 21d. The expression of Bcl-2 in treated group is higher than that in untreated group,and the statistics difference is significant (p<0.01) .
3 The positive expression of bFGF in normal group rat retina was mainly located at ganglion cell layer and inner nuclear layer.The staining in ganglion cell layer and inner nuclear layer were increased in untreated group ,and positive cells were observed in outer nuclear layer. The expression of bFGF in treated group is higher than that in untreated group,and the statistics difference is significant (p<0.01) .
4 Every sheet of cells in normal group rats retina lined up in order under the light microscope and TEM;Cells in untreated group lined up irregularly,the whole thickness thinningzed,and manifest apoptosis morphologic change was observed.Mohphologic change in treated group was lessened obviously.
Conclusions
1 0.2%brimonidine tartrate relieves the apoptosis morphologic change and elevate survival rate of RGCs, showing obvious effect in threapying rat optic nerve crush injury.
2 0.2% brimonidine tartrate elevates the expression of bFGF、Bcl-2 ,cuts down the expression of the Bax in optic nerve crush injury rat retina,this hints that the therapeutic mechanism of 0.2% brimonidine tartrate may relevants with its effects of promoting the expression of bFGF and inhibiting apoptosis.
引文
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33 周延冲, 主编. 多肽生长因子, 基础与临床[M ]. 北京: 中国科学技术出版社,1992. 209-232
34 Baird A,Boh len P.Fibroblast growth factors [M ]. In: Spo rn MB,Roberts AB, eds.Peptide growth factors and their receptors. New York: Sp ringer,1991.369-406
35 Hu DN,R itch R. Tissue culture of adult human retinal ganglion cells [J ]. J Glaucoma ,1997,6:37
36 Lipton SA,Wagner JA,Madison RD,et al.Acidic fibroblast growth factor enhances regeneration of processes bypostnatal mammalian retinal ganglion cells in culture[J ].Proc Natl Acad SciU SA,1988,85:2388-2392
37 周浩,周韵秋,魏锐利等.碱性成纤维细胞生长因子对兔急性高眼压视神经纤维损害的保护作用.中华眼底病杂志,2000,16(2):94-96
38 袁洪峰,刘少章,贺翔鸽.碱性成纤维生长因子在视神经中的表达及在视神经损伤后的反应.眼科研究,2002,20(2):132-134
39 Blanco RE, Lopez-Roca A, Soto J,et al. Basic fibroblast growth factor applied to the optic nerve after injuryincreases long term cell survival in the frogretina. J Comp Neurol,2000,423(4):646-658
40 Unok K,LaVail MM. Protection of the rat retina from ischemic injury by brain-derived neurotrophic factor, ciliary neurotrophic factor,and basic fibroblast growth factor[J ].Invest Ophthalmol Vis Sci, 1994,35:907-915
1 张宝恒,吴凤简.作用于 α 肾上腺素受体的药物.生理科学进展, 1990,21(2):178-186
2 Osborne NN. Binding of (-)[3H]noradrenaline to bovine membrane of the retina: evidence for the existence of alpha2-receptors. Vision Res.1982,22:1401-1407
3 Zarbin MA, Wamsley JK, Palacios JM, et al. Autoradiographic localization of high affinity GABA, benzodiazepine, dopaminergic, adrenergic and muscarinic cholinergic receptors in the rat, monkey and human retina. Brain Res. 1986,374:75-92
4 Wen R, Cheng T, Li T, et al. a2-Adrenergic agonists induced basic fibroblast growth factor expression in photoreceptors in vivo and ameliorate light damage. J Neurosci. 1996,16:5986-5992
5 Grundy JE,McKeating JA, Ward PJ, et al. Beta 2 microglobulin enhances the infectivity of cytomegalovirus and when bound to the virus enables class I HLA molecules to be used as a virus receptor. J Gen Virol 1987,68(pt3):793-803
6 Wheeler LA,Lai R, Woldemussie E. From the lab to the clinic: activ ation of an alpha-2 agonist pathway is neuroprotective in models of retinal and optic nerve injury. Eur J Ophthalmol, 1999; 9 (Suppl.1): S17-21
7 Thompson CD , Macdonald TL, Garst ME, et al. Mechanisms of adrenergic agonist induced allergy bioactivation and antigen form ation. Exp Eye Res,1997,64(5):767-773
8 LeBlanc RP. Twelve-month results of an ongoing randomized trial comparing brimonidine tartrate 0.2% and timolol 0.5% given twice daily in patients with glaucoma or ocular hypertension.Ophthalmol ogy,1998,105(10): 1960 -1967
9 Katz LJ. Brimonidine tartrate 0.2% twice daily vs timolol 0.5% twice daily:1-year results in glaucoma patients. Am J Ophthalmol, 1999,127:20-26
10 Enyr S.Arcieri , Rafael S.Arcieri,Ana C.A.Pereira et al.Comparing the fixed combination brimonidine-timolol versus fixed combinati on dorzolamide-timolol in patients with elevated intraocular pressur e.Current Medical Research and Opinion, 2007,23(4):683-689
11 孙丽,马海,刘磊等. 0.2%酒石酸溴莫尼定的降眼压作用.中国实用眼科杂志,2001,19(6):431-432
12 Toris CB,Camras CB, Yablonski ME. Acute versus chronic effects of brimonidine on aqueous humor dynamics in ocular hypertensive patients. Am J Ophthalmol,1999,128(1):8-14
13 Yoles E,Wheeler LA, Schwartz M. Alpha2-adrenoreceptor agonists are neuroprotective in a rat model of optic nerve degeneration. Inves t Ophthalmol Vis Sci,1999 ,40(1):65-73
14 Traverso CE. The Association for Research in Vision andOphthalm ology (ARVO) annual meeting. Invest Ophthal mol Vis Sci,1998,39 (4):S1-1134
15 Simmons ST. Association for Research in Vision and Ophthalmolog y annual meeting. Invest Ophthalmol Vis Sci, 2000,41(4):s1-959
16 Wang F.Association for Research in Vision and Ophthalmo logy 1996 annual meeting. Invest Ophthalmol Vis Sci,1996,37(3):S1-1154
17 Lee DA. Efficacy of brimonidine as replacement therapy in patients with open-angle glaucoma or ocular hypertension. Clin Ther,2000,22(1):53-65
18 Tomomi Metoki, Hiroshi Ohguro, Ikuyo Ohguro, et al. Study of effe cts of Antiglaucoma Eye Drops on N-Mithyl- D-Aspartate-Induced Retinal Damage. Jpn J Ophthalmol, 2005, 49:453-461
19 Kelly M. a2-Selective Adrenergic Agonists’s neuropro tective effect on retinal ganglion cells.Invest Ophthalmol Vis Sci, 1999; 40(4): 481-486
20 Dreyer EB. A proposed role for excitotoxicity in glaucoma. J Glauco ma,1998,7(1):62-67
21 Maria P.Lafuente,Maria P az Villegas-Perez,Paloma Sobrado -Calvo,et al. Neuroprotective Effects of a2-Select ive Adrenergic Agonists against Ischemia-Induced Retinal Ganglion Cell Death. Invest Ophthalmol Vis Sci,2001,42 (9):2074-2084
22 Yoles E,Wheeler LA,Schwartz M.Alpha2-adrenorecept oragonists are protective in a rat model of optic nerve deg eneration .Invest Ophthalmol Vis Sci,1999,40:65-73
23 Elizabeth WoldeMussie,Guadalupe Ruiz,Mercy Wijono, Neuroprote ction of Retinal Ganglion Cells by Brimonidine in Rats with Laser -Induced Chronic Ocular Hypertension . Invest OphthalmolVis Sci, 2001,42(12):2849-2855
24 Hasson D. Neuroprotective effect of alpha-2 agonist of ischemic rat retina. Soc Neurosci, 1997, 23(122):168-170
25 Ahmed F. Alteration of retinal intrinsic survival signal and effect of a2–adrenergic receptor agonist in the retina of the chronic ocular hypertension rat.Invest Ophthalmol Vis Sci,1999. 40(4): 265- 269
26 J. R. Ferencz,Gila Gilady,M. Belkin,et al. Topical brim onidine reduces collateral damage caused by laser photo coagulation for choroidal Neovascularization. Graefe’s Arch Clin Exp Ophthalmol, 2005,243:877–880
27 D W Ewans,S L Hosking,D Gherghel,et al.Contrast sensiti vity imp roves after brimonidine therapy in primary open angle glaucom a:a case for neuroprotection .Br J Ophthal mol,2003,87:1463-1465
28 李美玉.青光眼学,2004年8月第1版,人民卫生出版社,562-563
29 Zhang C , Takahashi K, Lam TT, et al. Effects of basic fibroblast growth factor in retinal ischemia.Invest Ophthal mol Vis Sci,1994,35(8): 3163-3168
30 Nancy S.Y.Yuen, Peggy Cheung, and Sui Ping Hui。Comparing Brim onidine 0.2% to Apraclonidine 1.0% in the Prevention of Intrao cular Pressure Elevation and Their Pupillary Effects Following Laser Per ipheral Iridotomy,Jpn J Ophthalmol 2005,49:89-92
31 Martinez CE,Applegate RA,Klyce SD,et al.Effect of papillarydila tion on corneal optical aberration after photo refractive keratectomy .Arch Ophthalmology 1998, 116 : 1053-1062
32 Kohnen T.The importance of reporting the complications of refracti ve surgery.J Cataract Refract Surg,1999,25:17-22
33 Thompson CD , Macdonald TL, Garst ME, et al. Mecha nisms of adrenergic agonist induced allergy bioactivation and antigen forma tion. Exp Eye Res,1997, 64(5):767-773
34 Haydar Erdogˇ an, M.I.lker Toker, M. Kemal Ar?c?,et al. Effect of Latanoprost 0.005% andBrimonidine 0.2% on Intraocular Pressure After Phacoemulsification and Intraoc ular Lens Implantation Surgery. Jpn J Ophthalmol, 2004,48: 598-601
35 Chevrier RL , Assalian A, Duperré J, et al. Apraclonidine 0.5% vers us brimonidine 0.2% for the control of intra ocular pressure elevation following anterior segment laser procedures. Ophthalmic Surg Las ers,1999,30 (3): 199-204
36 LeBlanc RP. Twelve-month results of an ongoing random ized trial comparing brimonidine tartrate 0.2% and timolol 0.5% given twice daily in patients with glaucoma or ocular hypertension. Ophthalmology,1998,105(10) :1960 -1967
37 Katz LJ. Brimonidine tartrate 0.2% twice daily vs timolol 0.5%twice daily: 1-year results in glaucoma patients. Am J Ophthalmol,1999,127:20-26
38 Melamed S ,David R. Ongoing clinical assessment of the safety pro file and efficacy of brimonidine compared with timolol: year-three results. Brimonidine Study Group II. Clin Ther. 2000,22(1): 103- 111.
39 Butler PJ.Effect of brimonidine on blood supply of normal rat retina .Invest Ophthalmol Vis Sci,1996,37(3):S201-213
40 Spada CS,Messmer K, Kubler W. Sixth World Congress for Microci rculation. 1996,5:11-15
41 Spada CS. Color Doppler ultrasonography in the evaluat ion oftwo kinds of eye drops on blood supply of visual nerve in glaucoma. Inv est Ophthalmol Vis Sci, 1995, 36: 1041-1047
42 Carlsson AM,Chauhan BC, Lee AA, et al. The effect of brimonidine tartrate on retinal blood flow in patients with ocular hypertension. Am J Ophthalmol,2000,129(3): 297- 301
2 Kallela I ,Hyrkas T ,Paukku P,et al.Blindness after maxillofacial blunttrauma. Evaluation of candidates for optic nerve decompression surgery.J ,Cranion axillofac Surg,1994,22(4):220-225
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4 Levin LA ,Beck RW,Joseph MP,et al.The treatmentof traumatic optic neuropathy:the International Optic N erve Trauma Study.J,Ophthalmology,1999,106(7):1268-1277
5 彭玉豪,李和平, Clark AF1.青光眼性视网膜神经节细胞凋亡的病理机制.中华眼科杂志,2004,40:495-4991
6 Berkelaar M,Clarke DB ,Wang YC. Axotomy results indelayed death and apoptosis of retinal ganglion cells in adult rats.J,Neurosci,1994,14(7):4368-4374.
7 Schumer RA.Arch Ophthalmol,1994,112:37
8 黄厚斌,张卯年,马志中.大鼠视神经不同程度量化损伤模型的建立和评价.中华创伤杂志,2004,20:747-750
9 Roth S ,Osinski JV,Park SS,et al.Measurement of purine nucleoside concentration in the intact rat reti na. J Neurosci Methods ,1996,68:87-90
10 Si-Wei You,Kwok-Fai So,Henry K.Yip .Axonal Regeneration of Retinal Ganglion Cells Depending on the Distance of Axotomy in Adult Hamsters.Invest Ophthalmol Vis Sci,2000,41:3165-3171
11 Hana Levkovitch-Verbin,Catby Harris-Cerruti,Yoram Groner.RGC Death in Mice after Optic Nerve Crush InjuryOxidative Stress and Neuroprotection. Invest Ophthalmol Vis Sci,2000,41:4169-4174
12 Maria P.Lafuente,Maria Paz Villegas-Perez,Paloma Sobrado-Calvo,et al. Neuroprotective Effects of a2-SelectiveAdrenergic Agonists against Ischemia-Induced Retinal Ganglion Cell Death. Invest Ophthalmol Vis Sci,2001,42(9):2074-2084
13 Yoles E,Wheeler LA,Schwartz M.Alpha2-adrenoreceptoragonists are protective in a rat model of optic nerve degeneration.Invest Ophthalmol Vis Sci,1999,40:65-73
14 Elizabeth WoldeMussie,Guadalupe Ruiz,Mercy Wijono, Neuroprotection of Retinal Ganglion Cells by Brimonidine in Rats with Laser-Induced Chronic Ocular Hypertension . Invest Ophthalmol Vis Sci,2001,42(12):2849-2855
15 Osborne NN. Binding of (-)[3H]noradrenaline to bovine membrane of the retina:evidence for the existenceofalpha2-receptors.VisionRes.1982,22:1401-1407
16 Zarbin MA, Wamsley JK, Palacios JM, et al.Autoradiographic localization of high affinity GABA, benzodiazepine, dopaminergic, adrenergic and muscarinic cholinergic receptors in the rat, monkey and human retina. Brain Res.1986,374:75-92
17 Wen R,Cheng T, Li T, et al.α2-Adrenergic agonists induced basic fibroblast growth factor expression in photoreceptors in vivo and ameliorate light damage. J Neurosci. 1996,16:5986-5992
18 Grundy JE.J Gen Virol 1987,60:793
19 Levin LA,Louhab A,Aref AB,et al.Apoptosis of retinalganglion cells in anerior ischemic optic neuropathy .J,Arch Ophthalmol,1996,114(4):488-491
20 Quigley HA,Nickells Rw,Kerrigan LA,et a1.Retinalganglion cell death in experimental glaucoma and afteraxotomy occurs by apoptosis.Invest Ophthalmol Vis Sci,1995,36:774-786
21 Chierzi S,Strettoi E,Cenni MC,et a1.Optic nerve crush:Axonal responses in wild-type and Bcl-2 transgenie mice.J Neurosei,1999,19:8367-8376
22 Motoyama N,Wang F,Roth KA,et al.Massive cell deathof immature hematopoietic cells and neurons in Bcl-2 deficient mice.J Science,1995,267(5203):1506-1510
23 Bonfanti L,Strettoi E,Chierzi S,et al.Protection of retinal ganglion cells from natural and axotomy-ind uced cell death in neonatal transgenic mice overexpre ssing Bcl-2.J Neurosci,1996,16(13):4186-4194
24 凌士奇,丁亚平,吕源叔.大鼠视网膜缺血再灌注损伤中的细胞凋亡.眼科,2003,12(1):47-49
25 傅强,侯铁胜,刘善荣等.大鼠脊髓急性损伤后Bcl-2和Bax的表达.中国矫形外科杂志,2001,8(3):259-260
26 Lou J ,Lenle LJ,Xu F,et al.In vivo Bcl-2 neuronal expression in the rat spinal cord.Science,1995,23(5):517-521
27 Porciatti V,Pizzorusso T,Cenni MC,et al.The visual response of retinal ganglion cells is not altered by optic nerve transection in trans-genic mice overexpressing Bcl-2. Proc Natl Acad Sci USA ,1996,93(25):141955-141959
28 Chierzi S,Cenni MC,Maffei L,et al.Protection of retinalganglion cells and preservation of function afteoptic nerve lesion in Bcl-2 transgenic mice.Vision Res,1998,38(10):1537-1543
29 Yin XM,Oltvai ZN,Korsmeyer SJ.BH1 and BH2 domains of Bcl-2 are requir-ed for inhibition of apoptosis and heterodimerization withBax.Nature,1994,369:312-313
30 吴永肯,刘彦文,邱呜新等.基因Bcl-2对神经元生物活性的保护作用.中国临床康复,2003,7(22):3071-3072
31 易少华,吴红色,邓伟连.视神经挫伤后视网膜形态学和Bcl-2/Bax表达.眼外伤职业眼病杂志,2006,28(5):321-324
32 牛建军,王一,孙时英.视神经钳夹伤后凋亡相关基因Bcl-2和Bax在大鼠视网膜上的表达.眼视光学杂志,2006,8(2):98-101
33 周延冲, 主编. 多肽生长因子, 基础与临床[M ]. 北京: 中国科学技术出版社,1992. 209-232
34 Baird A,Boh len P.Fibroblast growth factors [M ]. In: Spo rn MB,Roberts AB, eds.Peptide growth factors and their receptors. New York: Sp ringer,1991.369-406
35 Hu DN,R itch R. Tissue culture of adult human retinal ganglion cells [J ]. J Glaucoma ,1997,6:37
36 Lipton SA,Wagner JA,Madison RD,et al.Acidic fibroblast growth factor enhances regeneration of processes bypostnatal mammalian retinal ganglion cells in culture[J ].Proc Natl Acad SciU SA,1988,85:2388-2392
37 周浩,周韵秋,魏锐利等.碱性成纤维细胞生长因子对兔急性高眼压视神经纤维损害的保护作用.中华眼底病杂志,2000,16(2):94-96
38 袁洪峰,刘少章,贺翔鸽.碱性成纤维生长因子在视神经中的表达及在视神经损伤后的反应.眼科研究,2002,20(2):132-134
39 Blanco RE, Lopez-Roca A, Soto J,et al. Basic fibroblast growth factor applied to the optic nerve after injuryincreases long term cell survival in the frogretina. J Comp Neurol,2000,423(4):646-658
40 Unok K,LaVail MM. Protection of the rat retina from ischemic injury by brain-derived neurotrophic factor, ciliary neurotrophic factor,and basic fibroblast growth factor[J ].Invest Ophthalmol Vis Sci, 1994,35:907-915
1 张宝恒,吴凤简.作用于 α 肾上腺素受体的药物.生理科学进展, 1990,21(2):178-186
2 Osborne NN. Binding of (-)[3H]noradrenaline to bovine membrane of the retina: evidence for the existence of alpha2-receptors. Vision Res.1982,22:1401-1407
3 Zarbin MA, Wamsley JK, Palacios JM, et al. Autoradiographic localization of high affinity GABA, benzodiazepine, dopaminergic, adrenergic and muscarinic cholinergic receptors in the rat, monkey and human retina. Brain Res. 1986,374:75-92
4 Wen R, Cheng T, Li T, et al. a2-Adrenergic agonists induced basic fibroblast growth factor expression in photoreceptors in vivo and ameliorate light damage. J Neurosci. 1996,16:5986-5992
5 Grundy JE,McKeating JA, Ward PJ, et al. Beta 2 microglobulin enhances the infectivity of cytomegalovirus and when bound to the virus enables class I HLA molecules to be used as a virus receptor. J Gen Virol 1987,68(pt3):793-803
6 Wheeler LA,Lai R, Woldemussie E. From the lab to the clinic: activ ation of an alpha-2 agonist pathway is neuroprotective in models of retinal and optic nerve injury. Eur J Ophthalmol, 1999; 9 (Suppl.1): S17-21
7 Thompson CD , Macdonald TL, Garst ME, et al. Mechanisms of adrenergic agonist induced allergy bioactivation and antigen form ation. Exp Eye Res,1997,64(5):767-773
8 LeBlanc RP. Twelve-month results of an ongoing randomized trial comparing brimonidine tartrate 0.2% and timolol 0.5% given twice daily in patients with glaucoma or ocular hypertension.Ophthalmol ogy,1998,105(10): 1960 -1967
9 Katz LJ. Brimonidine tartrate 0.2% twice daily vs timolol 0.5% twice daily:1-year results in glaucoma patients. Am J Ophthalmol, 1999,127:20-26
10 Enyr S.Arcieri , Rafael S.Arcieri,Ana C.A.Pereira et al.Comparing the fixed combination brimonidine-timolol versus fixed combinati on dorzolamide-timolol in patients with elevated intraocular pressur e.Current Medical Research and Opinion, 2007,23(4):683-689
11 孙丽,马海,刘磊等. 0.2%酒石酸溴莫尼定的降眼压作用.中国实用眼科杂志,2001,19(6):431-432
12 Toris CB,Camras CB, Yablonski ME. Acute versus chronic effects of brimonidine on aqueous humor dynamics in ocular hypertensive patients. Am J Ophthalmol,1999,128(1):8-14
13 Yoles E,Wheeler LA, Schwartz M. Alpha2-adrenoreceptor agonists are neuroprotective in a rat model of optic nerve degeneration. Inves t Ophthalmol Vis Sci,1999 ,40(1):65-73
14 Traverso CE. The Association for Research in Vision andOphthalm ology (ARVO) annual meeting. Invest Ophthal mol Vis Sci,1998,39 (4):S1-1134
15 Simmons ST. Association for Research in Vision and Ophthalmolog y annual meeting. Invest Ophthalmol Vis Sci, 2000,41(4):s1-959
16 Wang F.Association for Research in Vision and Ophthalmo logy 1996 annual meeting. Invest Ophthalmol Vis Sci,1996,37(3):S1-1154
17 Lee DA. Efficacy of brimonidine as replacement therapy in patients with open-angle glaucoma or ocular hypertension. Clin Ther,2000,22(1):53-65
18 Tomomi Metoki, Hiroshi Ohguro, Ikuyo Ohguro, et al. Study of effe cts of Antiglaucoma Eye Drops on N-Mithyl- D-Aspartate-Induced Retinal Damage. Jpn J Ophthalmol, 2005, 49:453-461
19 Kelly M. a2-Selective Adrenergic Agonists’s neuropro tective effect on retinal ganglion cells.Invest Ophthalmol Vis Sci, 1999; 40(4): 481-486
20 Dreyer EB. A proposed role for excitotoxicity in glaucoma. J Glauco ma,1998,7(1):62-67
21 Maria P.Lafuente,Maria P az Villegas-Perez,Paloma Sobrado -Calvo,et al. Neuroprotective Effects of a2-Select ive Adrenergic Agonists against Ischemia-Induced Retinal Ganglion Cell Death. Invest Ophthalmol Vis Sci,2001,42 (9):2074-2084
22 Yoles E,Wheeler LA,Schwartz M.Alpha2-adrenorecept oragonists are protective in a rat model of optic nerve deg eneration .Invest Ophthalmol Vis Sci,1999,40:65-73
23 Elizabeth WoldeMussie,Guadalupe Ruiz,Mercy Wijono, Neuroprote ction of Retinal Ganglion Cells by Brimonidine in Rats with Laser -Induced Chronic Ocular Hypertension . Invest OphthalmolVis Sci, 2001,42(12):2849-2855
24 Hasson D. Neuroprotective effect of alpha-2 agonist of ischemic rat retina. Soc Neurosci, 1997, 23(122):168-170
25 Ahmed F. Alteration of retinal intrinsic survival signal and effect of a2–adrenergic receptor agonist in the retina of the chronic ocular hypertension rat.Invest Ophthalmol Vis Sci,1999. 40(4): 265- 269
26 J. R. Ferencz,Gila Gilady,M. Belkin,et al. Topical brim onidine reduces collateral damage caused by laser photo coagulation for choroidal Neovascularization. Graefe’s Arch Clin Exp Ophthalmol, 2005,243:877–880
27 D W Ewans,S L Hosking,D Gherghel,et al.Contrast sensiti vity imp roves after brimonidine therapy in primary open angle glaucom a:a case for neuroprotection .Br J Ophthal mol,2003,87:1463-1465
28 李美玉.青光眼学,2004年8月第1版,人民卫生出版社,562-563
29 Zhang C , Takahashi K, Lam TT, et al. Effects of basic fibroblast growth factor in retinal ischemia.Invest Ophthal mol Vis Sci,1994,35(8): 3163-3168
30 Nancy S.Y.Yuen, Peggy Cheung, and Sui Ping Hui。Comparing Brim onidine 0.2% to Apraclonidine 1.0% in the Prevention of Intrao cular Pressure Elevation and Their Pupillary Effects Following Laser Per ipheral Iridotomy,Jpn J Ophthalmol 2005,49:89-92
31 Martinez CE,Applegate RA,Klyce SD,et al.Effect of papillarydila tion on corneal optical aberration after photo refractive keratectomy .Arch Ophthalmology 1998, 116 : 1053-1062
32 Kohnen T.The importance of reporting the complications of refracti ve surgery.J Cataract Refract Surg,1999,25:17-22
33 Thompson CD , Macdonald TL, Garst ME, et al. Mecha nisms of adrenergic agonist induced allergy bioactivation and antigen forma tion. Exp Eye Res,1997, 64(5):767-773
34 Haydar Erdogˇ an, M.I.lker Toker, M. Kemal Ar?c?,et al. Effect of Latanoprost 0.005% andBrimonidine 0.2% on Intraocular Pressure After Phacoemulsification and Intraoc ular Lens Implantation Surgery. Jpn J Ophthalmol, 2004,48: 598-601
35 Chevrier RL , Assalian A, Duperré J, et al. Apraclonidine 0.5% vers us brimonidine 0.2% for the control of intra ocular pressure elevation following anterior segment laser procedures. Ophthalmic Surg Las ers,1999,30 (3): 199-204
36 LeBlanc RP. Twelve-month results of an ongoing random ized trial comparing brimonidine tartrate 0.2% and timolol 0.5% given twice daily in patients with glaucoma or ocular hypertension. Ophthalmology,1998,105(10) :1960 -1967
37 Katz LJ. Brimonidine tartrate 0.2% twice daily vs timolol 0.5%twice daily: 1-year results in glaucoma patients. Am J Ophthalmol,1999,127:20-26
38 Melamed S ,David R. Ongoing clinical assessment of the safety pro file and efficacy of brimonidine compared with timolol: year-three results. Brimonidine Study Group II. Clin Ther. 2000,22(1): 103- 111.
39 Butler PJ.Effect of brimonidine on blood supply of normal rat retina .Invest Ophthalmol Vis Sci,1996,37(3):S201-213
40 Spada CS,Messmer K, Kubler W. Sixth World Congress for Microci rculation. 1996,5:11-15
41 Spada CS. Color Doppler ultrasonography in the evaluat ion oftwo kinds of eye drops on blood supply of visual nerve in glaucoma. Inv est Ophthalmol Vis Sci, 1995, 36: 1041-1047
42 Carlsson AM,Chauhan BC, Lee AA, et al. The effect of brimonidine tartrate on retinal blood flow in patients with ocular hypertension. Am J Ophthalmol,2000,129(3): 297- 301